title: Visual SNP genotyping using asymmetric PCR and split DNA enzymes.
abstract: Harnessing and applying genomic technologies in resource limited environments demand a next generation of platforms, which are convenient, quick and easy to apply. We describe here a visual colour change assay that can be applied to SNP genotyping, which unlike traditional methods, does not adopt complicated procedures or expensive instrumentation, desirable features in bringing genetic capabilities outside the laboratory. Our strategy involved a two-step method that first enriched target genomic regions using asymmetric PCR, followed by direct in situ application of split DNA probes. In the presence of target sequences that perfectly matched the complementary probes, the split probes reassembled active DNAzymes, which catalysed a colour change reaction. A single-base mismatch (indicative of a polymorphism) prevented this reassembly and colour change, providing the means for accurate SNP calling. This is the first report, to our knowledge, that demonstrates successful visual SNP genotyping of actual human DNA samples using DNAzymes.
authors.affiliation: Defence Medical and Environmental Research Institute, DSO National Laboratories, 27 Medical Drive, Singapore 117510.
authors.forename: Jia Ling
authors.initials: JL
authors.lastname: Neo
authors.forename: Kanglie Darius
authors.initials: KD
authors.lastname: Aw
authors.forename: Mahesh
authors.initials: M
authors.lastname: Uttamchandani
mesh_terms: D000085:Acetates / Q000737:chemistry; D003116:Color; D004247:DNA / Q000032:analysis*; D015342:DNA Probes / Q000737:chemistry; D021881:DNA, Catalytic / Q000378:metabolism*; D015894:Genome, Human; D005838:Genotype; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D013053:Spectrophotometry / Q000379:methods
chemical_list: D000085:Acetates; D015342:DNA Probes; D021881:DNA, Catalytic; D004247:DNA; C018824:ammonium acetate